This work deals with monooxygenases (hydroxylases, epoxidases), arene oxide hydrases, and glutathione S-epoxide transferases of benzo(a)pyrene and analogs from mammals and tissue cultures. The solubilized monooxygenase system consists of a flavo-protein, a P-450 hemoprotein, and phospholipid or an iron-sulfur protein (adrenal mitochrondrial system). In connection with the purification of the components of the above enzyme and the study of their interaction with each other, substrates, inhibitors, and effectors (both small- and macro-molecules), we will optimize the application of aminimide-containing resins to affinity chromatography; we will extend the use of these resins to pellicular high-speed chromatography of biomolecules. Purified monooxygenase components will be studied by biochemical and other methods, such as electron paramagnetic resonance, circular dichroism, and fluorescence spectroscopies. Substrates, substrate analogs and inhibitors (and some of their spin-labeled derivatives) of monooxygenases, hydrases, and S-epoxide transferases will be synthesized and used to elucidate the reaction mechanism during catalysis and regulation in vitro, and, possibly, in vivo of these enzymes. Our objective is to elucidate in these enzyme systems the function of each protein component during catalysis, the stoichiometry, substrate specificity, reaction intermediates, and mechanism of oxygen-activation. Also, we want to affect the site and increase or decrease the rate of a specific reaction in vitro and in vivo. Such knowledge may be useful in understanding chemical carcinogenesis and possibly in controlling the harmful effects of polyhydrocarbon carcinogens, and in understanding steroid metabolism and steroid hormone action. The development of pellicular resins for ion-exchange, reverse-phase, and ligand-specific liquid high-speed chromatography of biomolecules will greatly expedite and enhance the resolution of purification techniques used in life sciences.